This review summarizes the recent developments in the application of electromigration techniques for the enantioselective analysis of drugs and metabolites in biological fluids. During the period covered by this review, it has been observed an increase in the use of negatively charged chiral selectors, particularly sulfobutyl ether-beta-cyclodextrin and sulfated-beta-cyclodextrin, and the combination of two different chiral selectors in the running buffer to obtain the resolution of drugs and their metabolites. Low detection limits as required for pharmacokinetic studies were obtained by using concentration techniques, including sample stacking procedures, and more sensitive detection systems, such as laser-induced fluorescence and mass spectrometry. Finally, the major points are discussed that can be considered to obtain reliable methods for enantioselective bioanalysis.